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罗琴, 谢会, 罗明, 李爱华, 黄琪, 龚成, 王雪, 李茂中. 22种呼吸道病原体核酸多重联检试剂盒检测效能评估[J]. 疾病监测, 2022, 37(1): 132-138. DOI: 10.3784/jbjc.202106100331
引用本文: 罗琴, 谢会, 罗明, 李爱华, 黄琪, 龚成, 王雪, 李茂中. 22种呼吸道病原体核酸多重联检试剂盒检测效能评估[J]. 疾病监测, 2022, 37(1): 132-138. DOI: 10.3784/jbjc.202106100331
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Luo Qin, Xie Hui, Luo Ming, Li Aihua, Huang Qi, Gong Cheng, Wang Xue, Li Maozhong. Evaluation of multiplex combined real-time PCR detection kit for nucleic acid detection of 22 respiratory pathogens[J]. Disease Surveillance, 2022, 37(1): 132-138. DOI: 10.3784/jbjc.202106100331
Citation: Luo Qin, Xie Hui, Luo Ming, Li Aihua, Huang Qi, Gong Cheng, Wang Xue, Li Maozhong. Evaluation of multiplex combined real-time PCR detection kit for nucleic acid detection of 22 respiratory pathogens[J]. Disease Surveillance, 2022, 37(1): 132-138. DOI: 10.3784/jbjc.202106100331
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22种呼吸道病原体核酸多重联检试剂盒检测效能评估

Evaluation of multiplex combined real-time PCR detection kit for nucleic acid detection of 22 respiratory pathogens

    摘要
  • 摘要:
      目的   通过对1种新型呼吸道病原体核酸多重联检试剂盒检测效能评价，为新型冠状病毒（SARS-CoV-2）、流行性感冒（流感）病毒等叠加流行的大规模检测提供适宜技术方法。
      方法   收集北京地区急性呼吸道感染病例阳性标本252份，采用4种核酸检测试剂盒（3种核酸多重检测试剂盒A、B、C及1种SARS-CoV-2检测试剂盒D）分别进行荧光聚合酶链式反应（PCR）检测，评价其与临床标本检测结果的一致性。 选取3份呼吸道病原体标本核酸，使用无核糖核酸（RNA）酶水10倍梯度稀释，并选取1份SARS-CoV-2阳性标本核酸稀释为10−1～10−6，评价A试剂盒的检出能力及批内重复性。
      结果   A、B、C试剂盒对甲型流感病毒、鼻病毒、腺病毒、肠道病毒、人偏肺病毒、副流感病毒3型、副流感病毒4型、冠状病毒NL63的检出率均为100%，Kappa值均为1，结果完全一致。 对乙型流感病毒、呼吸道合胞病毒、副流感病毒1型、副流感病毒2型、冠状病毒OC43、冠状病毒229E、冠状病毒HKU1及肺炎支原体的Kappa值范围为0.91～1。 3种核酸联检试剂盒对呼吸道病原体的最低检出限相当。 A与D试剂盒对SARS-CoV-2的阴阳性符合率均为100%，批内重复性显示在稀释度10−1～10−3范围内，2种试剂各重复检测5次变异系数（CV）值均<5%。 在稀释度10−4～10−5范围内，A试剂对ORF1ab基因检出率为40%，N基因检出率为80%，D试剂对ORF1ab基因检出率为30%，N基因检出率为20%。
      结论   A试剂盒与其他3种试剂盒对病毒载量较高的阳性标本检测的准确度无明显差异，检测性能相似；本研究为大规模核酸检测提供了检测试剂盒的多种选择。

     

    Abstract:
      Objective  To evaluate the detection consistency and power of a multiplex combined real-time PCR detection kits, and provide reference for the prevention and control of influenza plus SARS-CoV-2 infection.
      Methods  A total of 252 acute respiratory infectious samples were collected in Beijing. Four nucleic acid detection kits (A, B, C for multi nucleic acids and D for nucleic acid of SARS-CoV-2) were used for real time PCR, respectively, to evaluate the consistency of the test results. Three nucleic acid samples of respiratory pathogens in 10-fold dilutions with RNase-free water and 1 nucleic acid samples of SARS-CoV-2 in dilution of 10–1–10–6 folds were used to evaluate the detection power and intra-batch repeatability of kit A.
      Results  The positive coincidence rates of kit A, B and C to detect influenza A virus, rhinovirus, adenovirus, enterovirus, human metapneumovirus, parainfluenza virus type 3 and 4 and coronavirus NL63 were 100% (all Kappa values=1). The detection power of three assays for influenza B virus, respiratory syncytial virus, parainfluenza virus type 1 and 2, coronavirus OC43, 229E, HKU1 were similar, with the overall consistency rates of 99.17%–100.00% (all Kappa values ≥0.91). The detection power of kit A, B and C were similar (P>0.05). The SARS-CoV-2 negative and positive coincidence rates tested by kit A and kit D were 100%, the intra-batch repeatability showed the coefficient of variatiot (CV) of viral loads tested by these two kits were less than 5% in the dilution range of 10−1–10−3. In the dilution range of 10−4–10−5, the detection power of kit A for open reading frame (ORF)1ab gene was higher (40%) than kit D (30%), as well as for N gene (80% vs. 20%).
      Conclusion  There were no significant differences in the accuracy of the kit A and other three kits for positive samples with high viral loads, and the detection power were similar. This study provided a multiple choice for large-scale nucleic acid reference for the prevention and control of SARS-CoV-2 infection.

     

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